Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy
Using a confocal epi-illuminated microscope with a polarizing beam splitter and dual-channel detection of single-molecule fluorescence induced by pulsed laser excitation, a new application of the three-dimensional, real-time spectroscopic technique BIFL (burst integrated fluorescence lifetime) is in...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
1999
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| _version_ | 1826263465159819264 |
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| author | Schaflfer, J Volkmer, A Eggeling, C Subramaniam, V Striker, G Seidel, C |
| author_facet | Schaflfer, J Volkmer, A Eggeling, C Subramaniam, V Striker, G Seidel, C |
| author_sort | Schaflfer, J |
| collection | OXFORD |
| description | Using a confocal epi-illuminated microscope with a polarizing beam splitter and dual-channel detection of single-molecule fluorescence induced by pulsed laser excitation, a new application of the three-dimensional, real-time spectroscopic technique BIFL (burst integrated fluorescence lifetime) is introduced. BIFL allows simultaneous registration of fluorescence intensity, lifetime, and anisotropy. It is shown to be well-suited to identify the freely diffusing fluorescent molecule Rhodamine 123 and the Enhanced Yellow Fluorescent Protein via their characteristic fluorescence anisotropy using a time-resolved analysis. Furthermore, data analysis is discussed and rotational correlation times of single molecules are determined. Applications for multidimensional single-molecule identification are outlined. |
| first_indexed | 2024-03-06T19:52:12Z |
| format | Journal article |
| id | oxford-uuid:2454815d-e177-4739-878d-aed29224de7b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:52:12Z |
| publishDate | 1999 |
| record_format | dspace |
| spelling | oxford-uuid:2454815d-e177-4739-878d-aed29224de7b2022-03-26T11:49:27ZIdentification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence AnisotropyJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:2454815d-e177-4739-878d-aed29224de7bEnglishSymplectic Elements at Oxford1999Schaflfer, JVolkmer, AEggeling, CSubramaniam, VStriker, GSeidel, CUsing a confocal epi-illuminated microscope with a polarizing beam splitter and dual-channel detection of single-molecule fluorescence induced by pulsed laser excitation, a new application of the three-dimensional, real-time spectroscopic technique BIFL (burst integrated fluorescence lifetime) is introduced. BIFL allows simultaneous registration of fluorescence intensity, lifetime, and anisotropy. It is shown to be well-suited to identify the freely diffusing fluorescent molecule Rhodamine 123 and the Enhanced Yellow Fluorescent Protein via their characteristic fluorescence anisotropy using a time-resolved analysis. Furthermore, data analysis is discussed and rotational correlation times of single molecules are determined. Applications for multidimensional single-molecule identification are outlined. |
| spellingShingle | Schaflfer, J Volkmer, A Eggeling, C Subramaniam, V Striker, G Seidel, C Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy |
| title | Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy |
| title_full | Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy |
| title_fullStr | Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy |
| title_full_unstemmed | Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy |
| title_short | Identification of Single Molecules in Aqueous Solution by Time-Resolved Fluorescence Anisotropy |
| title_sort | identification of single molecules in aqueous solution by time resolved fluorescence anisotropy |
| work_keys_str_mv | AT schaflferj identificationofsinglemoleculesinaqueoussolutionbytimeresolvedfluorescenceanisotropy AT volkmera identificationofsinglemoleculesinaqueoussolutionbytimeresolvedfluorescenceanisotropy AT eggelingc identificationofsinglemoleculesinaqueoussolutionbytimeresolvedfluorescenceanisotropy AT subramaniamv identificationofsinglemoleculesinaqueoussolutionbytimeresolvedfluorescenceanisotropy AT strikerg identificationofsinglemoleculesinaqueoussolutionbytimeresolvedfluorescenceanisotropy AT seidelc identificationofsinglemoleculesinaqueoussolutionbytimeresolvedfluorescenceanisotropy |